Etiologic Risk Group (see link below): */Disease Information (sort of like the Intro to your Mini Research Write up): -most people infected are in rural areas in Western Africa who work outside and are exposed to the Tsetse fly which carries the protozoan - symptoms include swollen lymph nodes, fever, headaches, and can lead to paralysis, personality changes, confusion, poor coordination, and can lead to death - has some working drugs, but none that are effective without more severe side effects
**http://tritrypdb.org/tritrypdb/showRecord.do?name=GeneRecordClasses.GeneRecordClass&source_id=Tb927.1.710&project_id=TriTrypDB#Sequence**
*Target (protein/gene name): phosphoglycerate kinase
*NCBI Gene # or RefSeq#: 23858220
*Protein ID (NP or XP #) or Wolbachia#: XP_011771261.1
*Organism (including strain): trypanosoma brucei gambienese
Etiologic Risk Group (see link below): */Disease Information (sort of like the Intro to your Mini Research Write up):
-most people infected are in rural areas in Western Africa who work outside and are exposed to the Tsetse fly which carries the protozoan
- symptoms include swollen lymph nodes, fever, headaches, and can lead to paralysis, personality changes, confusion, poor coordination, and can lead to death
- has some working drugs, but none that are effective without more severe side effects
Link to TDR Targets page (if present): http://tdrtargets.org/targets/view?gene_name=Tb927.1.710
Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.):
**http://tritrypdb.org/tritrypdb/showRecord.do?name=SequenceRecordClasses.SequenceRecordClass&project_id=TriTrypDB&primary_key=Tb927_01_v5.1**
Essentiality of this protein: A lot of “significant loss of fitness in bloodstream forms”, some essential, some non-essential
Is it a monomer or multimer as biological unit? multimer
Complex of proteins?: No
Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism):
One finding Calmodulin along with calcium inhibits it in yeast:
http://www.sciencedirect.com/science/article/pii/S0167488904001843
One finding that shows when gene for PGK is stopped, it produces more of CAT reporter gene, which its enzyme can be toxic at high levels:
**http://www.sciencedirect.com/science/article/pii/S0166685106003252**
One finding showing PGK in group B sptreptococci (GBS) can be binded by plasminogen and actin and alters cytoskeleton of GBS:
**http://www.sciencedirect.com/science/article/pii/S0882401011001185**
bisphosphonate inhibitors of PGK in humans:
**http://www.sciencedirect.com/science/article/pii/S0960894X98000596**
*EC#: 2.7.2.3
Link to BRENDA EC# page: http://www.brenda-enzymes.org/enzyme.php?ecno=2.7.2.3&Suchword=&organism%5B%5D=Trypanosoma+brucei&show_tm=0
-- Show screenshot of BRENDA enzyme mechanism schematic:
Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):
-- link to Sigma (or other company) page for assay (see Sigma links below)
http://www.sigmaaldrich.com/content/dam/sigma-aldrich/docs/Sigma/Enzyme_Assay/3phosglyphoskinase.pdf
-- -or link (or citation) to paper that contains assay information
http://www.sciencedirect.com/science/article/pii/0166685181900839#
-- links to assay reagents (substrates) pages.
Magnesium Sulfate: http://www.sigmaaldrich.com/catalog/product/sial/m1880?lang=en®ion=US
ß-Nicotinamide Adenine Dinucleotide: http://www.sigmaaldrich.com/catalog/product/sigma/n8129?lang=en®ion=US
3-Phosphoglyceric Phosphokinase Enzyme Solution:
http://www.sigmaaldrich.com/catalog/product/sigma/p7634?lang=en®ion=US
--- List cost and quantity of substrate reagents, supplier, and catalog #
https://www.sigmaaldrich.com/catalog/product/sigma/p7634?lang=en®ion=US
Structure (PDB or Homology model):
**http://www.rcsb.org/pdb/explore/explore.do?pdbId=13pk**
-- PDB # or closest PDB entry if using homology model: 13PK
---- Query Coverage: 98%
---- Max % Identities: 93%
---- % Positives: 403/415(97%)
---- Chain used for homology: A
Current Inhibitors: actinomycin D, Sinefungin, SO4 2-, suramin, and Ib1
Expression Information (has it been expressed in bacterial cells):
In Fasciola hepatica:
http://www.sciencedirect.com/science/article/pii/0166685181900839#
Expression of Trypanosoma brucei in E. coli:
http://www.sciencedirect.com/science/article/pii/037811199090182Q
Purification Method:
Purification in E. coli:
http://www.sciencedirect.com/science/article/pii/037811199090182Q
Image of protein (PyMol with features delineated and shown separately):
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
1 mslkerksin ecdlkgkkvl irvdfnvpld dgnitndyri rsalpavqkv lteggscvlm
61 shlgrpkgvs maegkelrsa ggipgfeqka tlkpvakrls ellsrpvtfa pdclnaadvv
121 skmspgdvvl lenvrfykee gsksteerea makilasygd vyisdafgta hrdsatmtgi
181 pkilghgaag ylmekeisyf akvlgnpprp lvaivggakv sdkiqlldnm lqridyllig
241 gamaytflka qgysigismc eesklefars llkkaedrkv qiilpidhvc htefkavdsp
301 litedqnipe ghmaldigpk tiekyvqtig kcksaiwngp mgvfemvpys kgtfaiakam
361 grgthehglm siigggdsas aaelsgeakr mshvstggga slellegktl pgvtvldeke
*length of your protein in Amino Acids: 420 amino acids
Molecular Weight of your protein in kiloDaltons using the **Expasy ProtParam** website: 45231.3 kDa
Molar Extinction coefficient of your protein at 280 nm wavelength: 22265, with 0.492 Absorbance
TMpred graph Image:
*CDS Gene Sequence:
TTACTCCTTATCATCCAATACTGCAACACCGGGAAGCGTTTTGCCCTCGAGGAGTTCCAAAGACGCACCACCACCAGTTGAAACATGAGAAATACGTGCAGCTTCACCACAGAGTTCAGCCGCACCCGCACTCTC
GCCACCACCGATGATACTCATGAGTCCACGTTTCTGAGTTCCTCGACCCATGGCTTTCGCAATTGCAAATGTACCTTTGGAATAAGGAACCATTTCAAATACACCCATGGGACCGTTCCAAATGGCGCTCTTACACT
TCCCAATCGTCTGAACATATTTTTCAATAGTCTTGGGACCAATATCCAGAGCCATATGCCCTTCAGGGATGTTTTGATCCTCAGTTATCAATGGAGAATCCACAGCTTTGAATTCCGTGTGGCAAACATGATCAATTG
GAAGAATAATCTGCACCTTGCGGTCCTCCGCCTTCTTCAGCAGGGATCGAGCAAATTCAAGTTTACTTTCCTCGCACTTCGATTTTCCAATGCTGTAACCCTGAGCCTTCAGAAATGTGTATGCCATTGCACCACCA
ATTAAGAGATAATCGATGCGCTGCAACATGTTATCCAGAAGTTGGATCTTTTCGCTCACTTTCGCTCCACCAACGATAGCAACCAGCGGACGCGGCGGGTTACCAAGTACCTTAGCGAAGTATGAAATCTCCTTCT
CCATCAAATAACCGGCAGCACCGTGACCCAAAATCTTTGGAATTCCGGTCATGGTAGCACTGTCACGGTGAGCTGCACCAAAAGCATCACTGATGTAAACATCACCATATGACGCAAGGATCTTGGCCATGGCTTC
ACGTTCCTCAGCATTTTCACCGTTTTCTTCGCTGTAGAAGCGTACATTTTCAAGTAACATAACATTTCCACGCTTTAACCGCGAAACCTCTACTCTGGCATCCAAGCAGTCGTGAGCAAAATATACATGTTGGCCGA
GTAATTCGGCAAGCCTTACAGCAACTGGACGAAGACTGTATTGCTCTTCGTAGCCTGCACTAACACTCATCTGCGGTAACGCACTGTCTTTGTTTTCGTTTAAGTACTGCATAAGCAAAGATTTCTTCTCGTCCTCA
GGAAGGCGCGCAAACAGCATTGTTTTTCCAGATTTCACCTCAGGTGATATGTGCGTTGCCTCTTCCCGTGCACTGGATTTCGCTAATATTTTAACTTTCTCCTCTCCCGGTAGAACACTAAAAAACGCTGTTTTACC
TTTGTTGTCGTGAAACTGAGCCTCGTGATATCGCCGTCTTATATCTCGCACACCCGCAACCATTGAAACGCCTCTGGGCCGTCCAAGATGGCTCAGGAGAATACAAATACCACCCTGATCAATAACCCGTCGTATC
GTTGGAATCGCCGCACGGATTCGATAATCGTTAGTAATATGGCCATTTTGCATAGGAACGTTGAAATCGACACGCATAAGCACTCGCTTCGCCGTTAACGGCCAAACATCGCCCACAGACTTCTTCAGGCTTATGTT
GTTCTTCGCATTCGGTGCAGTGGACAT
*GC% Content for gene: 46.9%
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
CTGTTGTTGA TAATCCAGTA CTGTAATACT GGTAAGAGGT TCGCGCTGGA AGAATTCCAA CGGCGGACAA CGACGAGCTA AAATATGCGC AACACGTGCT CCTTTACAAC GGAATTCTCC AGAACCCGCA CGTTGGCAAC TACTGATGAC ACACACGAGT CGACGTTTTT ATCGTCTTCA ACCCACGGTT TTAGAAACTG TAAATGCACA TTCGGTATAA GAAATCACTT TAAGTACACT CACGGTACGG TCCCGAATGG GGCCTTAACT CTCCCAAACC GCCTCAACAT ATTCTTCAAC TCACTCGGAA CTAACATTCA GTCTCACATG CCCTTCCGGG ATGTCTTAAT TTTAAGTTAC CAGTGGCGCA TTCATAGCTT TGAGTTCCGG GTTGCAAACA TGATCAACTG GAAGAACAAC CTTCATCTTG CTGTTCTCAG GCTCTTGCAG CAAGGGAGCA GCAAATTCAA ATTTACATTC CTTGCCCTTC GGTTCTCCAA CGCCGTCACA CTGTCCCTTC AGAAGTGCGT TTGTCACTGT ACCACTAACT AAGAAATCAT TGATGCACTC CAACACGTTA TACAGAAGCT CGATTTATTT GCACACTTCC GCAGCACAAA CGACTCTAAC CAGCGAACAA GGCGAGTTAC GAAGTACCTT TCCGAAGTCT AGAATCTCTT GCTCCACCAG ATCACAGGAT CAACTGTGAC ACAAAACCTC TGGAACTCGG GTCACGGGTC AACTGTGACC GTATCATGCA CTAAATCGAT CACTGACGTC AACATTACGA TTTAGCGCAA AGACCTTGGT CATGGTTTCA CATTCCTGTC GATTTTCACG GTGTTCTTTG CTGTTGAGGC CTACATTTTT AAATAACACA ACATCTCCAC TCTTTAGCCG CGTAACTTGT ACAGCGGTAT TCAAGCGGTT GTGTCAAAGA TTTACATGCT GGCTGAGTAG TTCGGTAAAC CATATAGTAA TTGGACAAAA ACAGTCTTGC TCTTTGTGGC GTGCACCAAC ACACATTTGC GGTAGAGGAC CGTGTTTGTT TTTGTGTAAG TGCTTCATAA GCAACGATTC CTGCTTGTAT TGCGTAAGGC CCGCAAGCAA CACTGCTTTA GCCGTTTTCA CTTGCGGTGA TATGTTCGTT GCCTGTTCCC TTGCACTGGA TTCCGGTAGT ACTTTAACTT CCTGCTGAGC CGTTAGAATA CCAAAAAGCG GTGTTTCACC TTTGTTGTGG TGAAACTGAG TCTTGTTATT AGTCCCTCTT ACATAAGTCA TACCAGAAAC CACTGAAACG CCTCTGGCCC GAGCAAAATG GCACAAGAAA ATACGAACAC GACACTTATA AATAACCCGT CATACCGCTG GAATCGGCGT ACCGATAGCA TTATAGTGTC GAACATGGCG ATCCTTCATC GGAACGTGGA GATTGACACT CATAAGCACA GTCTCAGACG ATGACGACCA AACATTGCTC ACCGCCTTTT ACAGGCATAC GTTGTGTTAC GGATTAGGTG CTCAGGCCAT
*GC% Content for gene (codon optimized): 45.6%